摘要
目的建立恶性疟原虫pfcrt(Plasmodium falciparum chloroquine resistant transporter)等位基因多态性的单体型的研究疗法,比较我国海南省与东南亚及非洲地区恶性疟原虫的pfcrt等位基因多态性的单体型的特征。方法来自海南省19份恶性疟滤纸血样提取DNA,通过巢式PCR反应,扩增出包含第72～76、97位密码子的基因片段,根据限制性片段长度多态性(RFLP)分析结果,各选取6份突变型和野生型PCR产物进行DNA测序,检测第72～76、97位密码子序列,从而建立并分析我国海南省恶性疟prcrt等位基因的单体型。结果19份滤纸血样本提取的DNA全部扩增出1条195 bp的片段,酶切消化后,有11份出现1条100 bp左右的酶切片段,为野生型pfcrt等位基因,其余8份为1条195 bp的片段,为突变型。基因序列分析发现,野生型pfcrt等位基因的第72～76位密码子单体型为CVMNK,突变型为CVIET,第97位密码子未发现突变。结论我国海南省氯喹抗药性恶性疟原虫pfcrt等位基因第72～76位密码子的单体型与东南亚和非洲地区抗氯喹恶性疟原虫相应单体型一致。
Objective To develop a method for comparing the haplotypes of pfcrt polymorphism of Hainan Province with those from other areas of the world. Methods Nested PCR was used to amplify the polymorphic region including codon 72 to 76 and 97 of pfcrt gene. The PCR products were digested by ApoI restriction endonuclease to determine the allelic types. According to the allelic types, 6 products from each of mutant type and wild type were sequenced to analyze the haplotypes of pfcrt polymorphism. Results Bands in size of 195 bp appeared in all 19 samples. After Apol digestion, 11 samples contained one ApoI site when codon 76 of the pfcrt gene codes for lysine （K76）, which were visualized by the presertce of 98 bp and 97 bp restriction fragments, The DNA sequencing revealed that 6 samples of cloroquine resistant P. falciparum carried pfcrt alleles encoding an amino acid haplotype of CVIET （residues 72-76）, and the haplotype of CVMNK was found in other 6 samples with wild type pfcrt gene. Conclusion Haplotypes of pfcrt polymorphism from Hainan were the same as those from southeast Asia and Africa.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2007年第2期106-108,113,共4页
Chinese Journal of Parasitology and Parasitic Diseases
